Journal article
Immunization of MERS-CoV-2-infected mice with a sublethal dose of MERS-CoV or VRP-MERS-S
STAR protocols, Vol.4(2), 102171
02/27/2023
DOI: 10.1016/j.xpro.2023.102171
PMCID: PMC9968614
PMID: 36920912
Abstract
Here, we detail the immunization of mice with a sublethal dose of MERS-CoV or two doses of replication-incompetent alphavirus replicon particles expressing MERS-CoV spike protein. We then describe steps to determine the outcome of immunization by challenging immunized mice with a lethal dose of MERS-CoV, as well as by detecting virus-specific neutralizing antibody and virus-specific T cell response via neutralization assay and flow cytometry, respectively. This protocol can be used to evaluate other CoV infections or vaccine-induced immune responses.
For complete details on the use and execution of this protocol, please refer to Zheng et al. (2021).1
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•Mouse model to determine MERS-CoV-specific immune responses•A replication-incompetent alphavirus VRP-MERS-S is produced to immunize mouse model•Protective effects induced by immunization are evaluated via in vivo and ex vivo systems•Virus-specific T cells are determined by flow cytometry following peptide stimulation
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Here, we detail the immunization of mice with a sublethal dose of MERS-CoV or two doses of replication-incompetent alphavirus replicon particles expressing MERS-CoV spike protein. We then describe steps to determine the outcome of immunization by challenging immunized mice with a lethal dose of MERS-CoV, as well as by detecting virus-specific neutralizing antibody and virus-specific T cell response via neutralization assay and flow cytometry, respectively. This protocol can be used to evaluate other CoV infections or vaccine-induced immune responses.
Details
- Title: Subtitle
- Immunization of MERS-CoV-2-infected mice with a sublethal dose of MERS-CoV or VRP-MERS-S
- Creators
- Jian Zheng - University of LouisvilleRudra Channapannavar - Oklahoma State UniversityStanley Perlman - Departments of Microbiology and Immunology, University of Iowa, Iowa City, IA 52242, USA
- Resource Type
- Journal article
- Publication Details
- STAR protocols, Vol.4(2), 102171
- DOI
- 10.1016/j.xpro.2023.102171
- PMID
- 36920912
- PMCID
- PMC9968614
- NLM abbreviation
- STAR Protoc
- ISSN
- 2666-1667
- eISSN
- 2666-1667
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 02/27/2023
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Infectious Disease (Pediatrics)
- Record Identifier
- 9984375454902771
Metrics
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